DocumentCode :
1231248
Title :
An On-Chip Fluorogenic Enzyme Assay Using a Multilayer Microchip Interconnected With a Nanocapillary Array Membrane
Author :
Gong, Maojun ; Kim, Bo Young ; Flachsbart, Bruce R. ; Shannon, Mark A. ; Bohn, Paul W. ; Sweedler, Jonathan V.
Author_Institution :
Dept. of Chem., Univ. of Illinois at Urbana-Champaign, Urbana, IL
Volume :
8
Issue :
5
fYear :
2008
fDate :
5/1/2008 12:00:00 AM
Firstpage :
601
Lastpage :
607
Abstract :
Microfluidic devices allow manipulation of reagents and fluids in a semi-automated fashion, ideal for performing multiple measurements or conditioning various reagents. Here, an enzyme assay has been performed in a multilayer poly(methyl methacrylate)-based microfluidic device, where the layers are fluidically connected via a nanocapillary array membrane serving as an effective injector and valve. As a model system, beta-glucuronidase from Escherichia coli and fluorescein di(beta-D-glucuronide) are used for the assay; offline mixing and online incubation of substrate and enzyme allow determination of the initial hydrolysis rates of the substrate under catalysis by beta-glucuronidase. The Michaelis constant Km was determined to be ~4.0 muM for the enzyme of 83 units/mL at ambient temperature. The 50% inhibitory concentration IC50 of D-saccharic acid-1,4-lactone to 167 units/mL was estimated to be 3.0 muM. These results demonstrate added functionality for a poly(methyl methacrylate)-based nanocapillary array membrane-containing microfluidic device for following enzyme reaction kinetics.
Keywords :
bioMEMS; capillarity; catalysis; enzymes; membranes; microfluidics; microprocessor chips; reaction kinetics; Escherichia coli; beta-glucuronidase; enzyme reaction kinetics; fluorescein di(beta-D-glucuronide); hydrolysis rates; microfluidic devices; multilayer microchip; nanocapillary array membrane; on-chip fluorogenic enzyme assay; Biochemistry; Biomembranes; Chemistry; Electrokinetics; Kinetic theory; Microfluidics; Nanoscale devices; Nonhomogeneous media; Performance evaluation; Valves; Enzyme kinetics; microcontact printing; microfluidics; nanocapillary array membranes; nanofluidics; poly(methyl methacrylate);
fLanguage :
English
Journal_Title :
Sensors Journal, IEEE
Publisher :
ieee
ISSN :
1530-437X
Type :
jour
DOI :
10.1109/JSEN.2008.918912
Filename :
4529140
Link To Document :
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